人胚胎干细胞培养及其cDNA文库的构建

目的构建人胚胎干细胞cDNA文库,为筛查疾病抗原基因奠定基础。方法培养和收集人胚胎干细胞克隆,提取胚胎干细胞总RNA,分离纯化mRNA,利用mRNA作模板反转录合成双链cDNA,双链DNA经pfu-DNA聚合酶将链末端补平,与EcoRⅠ接头连接,XhoⅠ酶切消化产生粘端。用Sepharose CL2B柱分离纯化及除去小分子cDNA片段,与Uni-ZAP XR噬菌体连接,体外转化XL1-blue MRF'菌,建成cDNA文库并计算重组效率。对cDNA文库进行扩增,测定扩增文库的滴度。阳性重组子用EcoRⅠ+XhoⅠ双酶切鉴定插入片段的大小。结果构建成含1.2×106重组子的人胚胎干细胞cDNA文库,重组子插入外源DNA片段长度不小于1000kb。结论已成功构建人胚胎干细胞cDNA文库,适合用于筛选目的基因克隆。

Anti-inflammatory effects of bifidobacteria by inhibition of LPS-induced NF-κB activation

AIM: Different strains of bifidobacteria were analysed for their effects on HT-29 intestinal epithelial cells (IECs) in in vitro models both of the non-inflamed and inflamed intestinal epithelium. METHODS: A reporter gene system in HT-29 cells was used to measure levels of NF-KB activation after challenge with bifidobacteria or after bacterial pre-treatment following LPS challenge. IL-8 protein and pro-inflammatory gene expression was investigated using normal HT-29 cells. RESULTS: None of the bifidobacteria tested induced activation of nuclear factor KB (NF-KB) indicating that bifidobacteria themselves do not induce inflammatory events in IECs. However, six out of eight bifidobacteria tested inhibited lipopolysaccharide- (LPS-) induced NFKB activation in a dose- and strain-dependent manner. In contrast, NF-KB activation in response to challenge with tumor necrosis factor-α(TNF-α) was affected by none of the tested bifidobacteria, indicating that the inhibitory effect of bifidobacteria is specific for LPS-induced inflammation in IECs. As shown with two of the six inhibitionpositive bifidobacteria, LPS-induced inhibition of NFKB activation was accompanied by a dose-dependent decrease of interleukin 8 (IL-8) secretion and by lower mRNA levels for IL-8, TNF-a, cyclooxygenase 2 (Cox-2), and intercellular adhesion molecule 1 (ICAM-1). CONCLUSION: Some strains of bifidobacteria are effective in inhibiting LPS-induced inflammation and thus might be appropriate candidates for probiotic intervention in chronic intestinal inflammation.

Is autoimmune hepatitis a frequent finding among HCV patients with intense interface hepatitis?

AIM:To evaluate the overlap of autoimmune hepatitis in hepatitis C virus(HCV)-infected patients with intense interface hepatitis.METHODS:Among 1759 patients with hepatitis C submitted to liver biopsy,92(5.2%) presented intense interface hepatitis.These patients were evaluated regarding the presence of antinuclear antibody(ANA),anti-smooth muscle antibody(SMA) and anti-liver/kidney microsomal antibody(LKM-1),levels of γ-globulin and histological findings related to autoimmune hepatitis(plasma cell infiltrate and presence of rosettes).RESULTS:Among patients with hepatitis C and intense interface hepatitis there was a low prevalence of autoantibodies(ANA=12%,SMA=5%,LKM-1=0%) and the median γ-globulin level was within the normal range.Typical histological findings of autoimmune disease were observed in only two cases(2%).After applying the score for diagnosis of autoimmune hepatitis,only one patient was classified with a definitive diagnosis of autoimmune hepatitis.Since overlap with autoimmune hepatitis was not the explanation for the intense necroinflammatory activity in patients with chronic hepatitis C we sought to identify the variables associated with this finding.The presence of intense interface hepatitis was associated with more advanced age,both at the time of infection and at the time of the biopsy,and higher prevalence of blood transfusion and alcohol abuse.CONCLUSION:Although possible,overlap with autoimmune hepatitis is a very rare association in HCV-infected patients with intense interface hepatitis,an unusual presentation which seems to be related to other host variables.

Hematopoietic cell transplantation for Crohn’s disease;is it time?

AIM: To review all studies in the literature that have assessed Hematopoietic cell transplantation (HCT) and Crohn’s disease (CD) with the ultimate aims of determining if this is a viable treatment option for those with CD. A secondary aim was to review the above literature and determine if the studies shed further light on the mechanisms involved in the pathogenesis of CD. METHODS: An extensive Medline search was performed on all articles from 1970 to 2005 using the ; bone marrow transplant, stem cell, hematopoietic cell, Crohn’s disease and inflammatory bowel disease. RESULTS: We identified one case in which a patient developed CD following an allogeneic HCT from a sibling suffering with CD. Evidence for transfer of the genetic predisposition to develop CD was also identified with report of a patient that developed severe CD following an allogeneic HCT. Following HCT it was found that the donor (that had no signs or symptoms of CD) and the recipient had several haplotype mismatches in HLA class Ⅲ genes in the IBD3 locus including a polymorphism of NOD2/CARD15 that has been associated with CD. Thirty three published cases of patients with CD who underwent either autologous or allogeneic HCT were identified. At the time of publication 29 of these 33 patients were considered to be in remission. The median follow-up time was seven years, and twenty months for allogeneic and autologous HCT respectively.For patients who underwent HCT primarily for treatment of their CD there have been no mortalities related to transplant complications. CONCLUSION: Overall these preliminary data suggest that both allogeneic and autologous HCT may be effective in inducing remission in refractory CD. Thissupports the hypothesis that the hematolymphatic cells play a key role in CD and that resetting of the immune system may be a critical approach in the management or cure of CD.

Diabetes mellitus, insulin resistance and hepatitis C virus infection: A contemporary review

AIM To summarise the literature data on hepatitis C virus(HCV)-infected patients concerning the prevalence of glucose abnormalities and associated risk.METHODS We conducted a PubM ed search and selected all studies found with the key words "HCV" or "hepatitis C virus" and "diabetes" or "insulin resistance". We included only comparative studies written in English or in French, published from January 2000 to April 2015. We collected the literature data on HCV-infected patients concerning the prevalence of glucose abnormalities [diabetes mellitus (DM) and insulin resistance (IR)] and associated risk [i.e., severe liver fibrosis, response to antivirals, and the occurrence of hepatocellular carcinoma(HCC)]. RESULTS HCV infection is significantly associated with DM/IR compared with healthy volunteers and patients with hepatitis B virus infection. Glucose abnormalities were associated with advanced liver fibrosis, lack of sustained virologic response to interferon alfa-based treatment and with a higher risk of HCC development. As new antiviral therapies may offer a cure for HCV infection, such data should be taken into account, from a therapeutic and preventive point of view, for liver and non-liver consequences of HCV disease. The efficacy of antidiabetic treatment in improving the response toantiviral treatment and in decreasing the risk of HCC has been reported by some studies but not by others. Thus, the effects of glucose abnormalities correction in reducing liver events need further studies.CONCLUSION Glucose abnormalities are strongly associated with HCV infection and show a negative impact on the main liver related outcomes.

Evaluation of the Immunoregulatory Capacities of Feed Microbial Materials in Porcine Intestinal Immune and Epithelial Cells

The establishment of drug-free feeding systems has been required for secure and healthy livestock production. Although functional feed materials containing microorganisms as alternatives to enhance intestinal immunity are expected to be beneficial for reducing diarrhoea caused by pathogens in weaned piglets, the effects of such materials on porcine intestinal cells have not been investigated in detail. Therefore, this work evaluated the immunoregulatory functions of microbial feed materials in porcine intestinal immune and epithelial cells. Porcine immune cells isolated from Peyer’s patches and mesenteric lymph nodes were stimulated with six different feed materials containing microorganisms, and evaluated for lymphocyte mitogenicity and cytokine inductions. In addition, porcine intestinal epithelial cells were stimulated with the materials before treatment with heat-killed enterotoxigenic Escherichia coli (ETEC), and analyzed for the proinflammatory cytokine expressions. The material containing Bifidobacterium thermophilum significantly augmented lymphocytes’ mitogenicity and also induced a high expression of IL-2, IL-6 and IFN-γ in immune cells, and inhibited ETEC-induced overexpression of IL-6 and IL-8 via regulation of Toll-like receptor signaling. These results suggest that this feed material stimulates intestinal epithelial and immune cells to exert immunoregulation, suggesting that this feed is expected to contribute to promoting the health of piglets without using antimicrobial feed materials.

Prion Protein Binds to Aldolase A Produced by Bovine Intestinal M Cells

Microfold (M) cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer’s patches. They can transport antigens and microorganisms to lymphoid tissues. Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disorder in cattle. It is linked to variant Creutzfeldt-Jakob disease in humans. Although it is thought that M cells transport the BSE agent, the exact mechanism by which it crosses the intestinal barrier is not clear. We have bovine intestinal epithelial cell line (BIE cells), which can differentiate into the M cell type in vitro after stimulation, and which is able to transport the BSE agent. We show here that M cells are able to incorporate large numbers of PrP coated magnetic particles into intracellular vesicles, which we collected. The results of 2-DE show a specific protein associated with the PrP-coated particles, compared with non-coated particles. This protein was identified as aldolase A, a glycolytic pathway enzyme, using LC-MS/MS analysis. Aldolase A was synthesized and secreted by BIE cells, and increased during M cell differentiation. In the villi of the bovine intestine, aldolase A was detected on the surface of the epithelium and in the mucus droplet of goblet cells. In the FAE of bovine jejunal and ileal Peyer’s patches, aldolase A was localized on the surface and the apical part of the M cells. The binding of rbPrP to aldolase A was clearly detected and inhibited by pre-treatment of anti-aldolase A antibody. Aldolase A was co-stained with incorporated PrPSc in M-BIE cells. These results suggest that bovine M cells and goblet cells synthesize aldolase A, and that aldolase A may have the ability to bind PrP and associate with PrP in cellular vesicles. Therefore, aldolase A-positive M cells may play a key role in the invasion of BSE into the body.

重度特应性皮炎与产生IL-10的变应原特异性CD4^+ T细胞的频率下降有关

Background.Several studies have investigated levels of T-cell-derived interleukin(IL)-10 in individuals with atopic dermatitis,with conflicting results.Aims/Hypothesis.In order to address whether stratification of disease severity may help resolve the different findings,the hypothesis was tested that individuals with severe atopic dermatitis have a lower frequency of circulating IL-10-producing,allergen-specific CD4+T cells than do individuals with mild disease.Methods.Using peripheral blood mononuclear cells derived from individuals with severe(n = 12)and mild atopic dermatitis(n = 10)and from nonatopic controls(n = 10),we investigated production by CD4+T cells of tumour necrosis factor(TNF)-α,IL-4,IL-5,IL-13 and IL-10 in response to phorbol myristate acetate/ionomycinand Der p1 allergen.Results.It was observed that there were significantly higher frequencies of allergen-specific circulating CD4+T cells producing TNF-α-IL-4-,IL-5-and IL-13,and lower frequencies of these cells producing IL-10 in individuals with severe atopic dermatitis compared with mildly affected individuals and nonatopic controls(P < 0.01 for all comparisons).Furthermore,the Der p1-specific CD4+T cells were enriched within the subset of cells positive for cutaneous lymphocyte-associated antigen.Conclusions.Analysis of levels of allergen-specific CD4+T-cell production of IL-10 in relation to disease severity argues in favour of a role for IL-10 in the control of atopic dermatitis.

Molecular Determinants Responsible for the Subcellular Localization of HSV-1 UL4 Protein

The function of the herpes simplex virus type 1 (HSV-1) UL4 protein is still elusive.Our objective is to investigate the subcellular transport mechanism of the UL4 protein.In this study,fluorescence microscopy was employed to investigate the subcellular localization of UL4 and characterize the transport mechanism in living cells.By constructing a series of deletion mutants fused with enhanced yellow fluorescent protein (EYFP),the nuclear export signals (NES) of UL4 were for the first time mapped to amino acid residues 178 to 186.In addition,the N-terminal 19 amino acids are identified to be required for the granule-like cytoplasmic pattern of UL4.Furthermore,the UL4 protein was demonstrated to be exported to the cytoplasm through the NES in a chromosomal region maintenance 1 (CRM1)-dependent manner involving RanGTP hydrolysis.

Bifidobacteria Upregulate Expression of Toll-Like Receptor Negative Regulators Counteracting Enterotoxigenic <i>Escherichia coli</i>Mediated Inflammation in Bovine Intestinal Epitheliocytes

We previously established a bovine intestinal epithelial cell line (BIE cells) and showed that BIE cells are useful in vitro model system for the study of interactions between pathogenic and beneficial microorganisms and bovine intestinal epithelial cells (IECs). In the present study, we aimed to select potential immunomodulatory bifidobacteria that may be used to beneficially modulate the inflammatory response in bovine IECs. We also aimed to gain insight into the molecular mechanisms involved in the anti-inflammatory effect of bifidobacteria by evaluating the role of Toll-like receptor (TLR)-2 and TLR negative regulators in the regulation of proinflammatory cytokines production and MAPK, NF-κB and PI3K pathways activation in BIE cells. Five bifidobacteria strains were evaluated in this study and according to their capacity to modulate the inflammatory response of BIE cells. Despite the unique effect of each strain, four common points were found when comparing the effect of the high and moderate anti-inflammatory strains: 1) Upregulation of TLR negative regulators and the intensity of that upregulation was related to the different immunomodulatory capacity of each bifidobacteria strain;2) The balance between MAPK activation and MKP-1 upregulation affected the anti-inflammatory effect of bifidobacteria in BIE cells;3) The inhibition of PI3K pathway was related to the anti-inflammatory effect of bifidobacteria;4) The immunoregulatory effect of bifidobacteria in BIE cells is partially dependent on TLR2. This study shows that BIE cells can be used for the selection of immunoregulatory bifidobacteria and for studying the mechanisms involved in the protective activity of immunobiotics against TLR4-induced inflammatory damage. In addition, we have demonstrated that the anti-inflammatory effect of bifidobacteria was achieved by a complex interaction of multiple TLRs negative regulators as well as the inhibition/activation of multiple signaling pathways.

Quantitative Microbiological Evaluation of <i>Salmonella</i>Typhimurium Shed in Diarrhea, Loose, and Normal Stools of Infected Pigs

Control of within-herd transmission of Salmonella is important for reducing the prevalence of this organism on pig farms and for preventing Salmonella-contamination of pork. At the farm level, understanding the within-herd transmission of Salmonella can lead to more effective control. Salmonella infection is dependent on the inoculation dose;hence, quantitative evaluation of Salmonella shed in feces would provide useful information for developing effective measures. In this study, to reproduce and evaluate the number of Salmonella shed in diarrhea, loose stools, and normal feces, weaned pigs were inoculated with 3.2 × 109, 3.2 × 107, and 3.2 × 105 cfu of Salmonella Typhimurium, respectively. The number of S. Typhimurium shed in the feces peaked within 1 week post-inoculation in every group and the most amount of diarrhea and loose stools were observed within 2 weeks post-inoculation. Diarrhea occurred 10 times (six pigs), and loose stools were observed 25 times (11 pigs). The average concentration of S. Typhimurium shed in diarrhea, loose stools, and normal feces was 1.0 × 108, 1.6 × 104, and 7.1 × 101 cfu/g feces, respectively. These data suggest that diarrhea and loose stools are significant sources of within-herd transmission of Salmonella. Moreover, as some of the normal feces contained >1.0 × 106 cfu/g of S. Typhimurium, even normal feces could be a source of within-herd transmission of Salmonella. At Salmonella-positive farms, reduction of the amount of Salmonella shed even in normal feces would lead to better control of within-herd transmission of Salmonella. These data can contribute to the control of within-herd transmission of Salmonella, particularly during the weaning period.

MicroPath-A pathway-based pipeline for the comparison of multiple gene expression profiles to identify common biological signatures

High throughput gene expression analysis is swiftly becoming the focal point for deciphering molecular mechanisms underlying various dif-ferent biological questions. Testament to this is the fact that vast volumes of expression profiles are being generated rapidly by scientists worldwide and subsequently stored in publicly available data repositories such as ArrayEx-press and the Gene Expression Omnibus (GEO). Such wealth of biological data has motivated biologists to compare expression profiles gen-erated from biologically-related microarray ex-periments in order to unravel biological mecha-nisms underlying various states of diseases. However, without the availability of appropriate software and tools, they are compelled to use manual or labour-intensive methods of com-parisons. A scrutiny of current literature makes it apparent that there is a soaring need for such bioinformatics tools that cater for the multiple analyses of expression profiles. In order to contribute towards this need, we have developed an efficient software pipeline for the analysis of multiple gene expression data-sets, called Micropath, which implements three principal functions;1) it searches for common genes amongst n number of datasets using a number crunching method of comparison as well as applying the principle of permutations and combinations in the form of a search strat-egy, 2) it extracts gene expression patterns both graphically and statistically, and 3) it streams co-expressed genes to all molecular pathways belonging to KEGG in a live fashion. We sub-jected MicroPath to several expression datasets generated from our tolerance-related in-house microarray experiments as well as published data and identified a set of 31 candidate genes that were found to be co-expressed across all interesting datasets. Pathway analysis revealed their putative roles in regulating immune toler-ance. MicroPath is freely available to download from: www.1066technologies.co.uk/micropath.

Rejection of Experimental Hodgkins Lymphoma by T-Cells Engineered with a CD19 Chimeric Antigen Receptor

T cells engineered to express chimeric antigen receptors (CARs) combining an external antibody binding domain with the CD3ζ T cell receptor (TCR) signaling domain for triggering cell activation are being used for immunotherapeutic targeting of tumor cells in a non-HLA restricted manner. In this study we transduced T cells with a CD19-CAR construct containing a truncated CD34 gene (tCD34) marker and used these to target the B cell antigen CD19 on the surface of a Hodgkin’s lymphoma (HL) cell line (L591) both in vitro and in vivo. Levels of tCD34 expression in transduced peripheral blood mononuclear cells (PBMCs) ranged from 6% - 20% and this was increased to 82% after selection for transduced tCD34+ cells. In vitro cytotoxicity testing on a CD19+ HL cell line (L591) showed specific cell lysis initiated by the CD19-CAR transduced PBMCs. Importantly, CD19-CAR T cells prevented the growth of L591 HL tumor cells when co-injected subcutaneously (sc) in 6/6 severe combined immunodeficient (SCID) mice. There was no evidence of anti-tumor activity when CD19-CAR T cells were infused intravenously (iv) at the same time as L591 HL tumor cells were injected sc. However, 3/6 SCID mice showed tumor rejection within 83 days after iv infusion of CD19-CAR T cells 3 - 9 days after establishment of L591 HL tumors, while all control animals succumbed to tumors within 60 days. Interestingly, immuno-histochemical analysis of L591 HL tumors demonstrated that CD19-CAR T cells were detected not earlier than 11 days after infusion within the tumor mass. These results suggest that CD19 is a potentially attractive target for the immunotherapy of HL.

Xenobiotics-Induced Liver Damage Is Biochemically Abrogated by Treatment with Lipophilic Extracts of <i>Moringa oleifera in Vivo</i>

Context Drug-induced hepatotoxicity represents a significant proportion of liver disease cases. Currently, there is no effective treatment. To date efforts to identify treatment regimen that can reverse progressive damage have not been successful. We have previously shown that extract from Moringa (M) oleifera possesses clinically relevant antidiabetic and electrolyte modulators. Objective The aim of the current studies is to create experimental model of xenobiotic induced liver damage and investigate if treatment with lipophilic extract of M. oleifera could biochemically reverse progressive liver damage. Materials and Method For two groups of healthy rats, 7 in each group received 200 mg of extract or vehicle twice daily for 14 days. Acute toxicity, hepatotoxicity and hematologic/endothelial toxicity were monitored. Then 30 rats weighing 130 - 200 g received repeated dose of acetaminophen (xenobiotics) (640 mg/kg) for 5 days. Hepatotoxicity was confirmed biochemically by an established protocol. Treatment with M. oleifera extract resulted in mean weight of 132.2 ± 5.05 compared to the control with 134.1 ± 5.08 (P > 0.8115) among the healthy rats. Their LDH levels were 170.7 ± 13.02 and 133.8 ± 7.17 (P > 0.0698) for controls group, while the mean serum (ALT) level was 12.4 ± 1.2 or 25.6 ± 5.644 (P M. oleifera resulted in 100% biochemical recovery from hepatitis compared to the control group (P M. oleifera could effectively and biochemically abrogate xenobiotics induced liver damage in animal model.

Alterations of Blood Immune Cells in Elderly Patients with COVID-19 and with or without Pre-Existing Type 2 Diabetes

Elderly individuals, especially those with pre-existing conditions like type 2 diabetes mellitus (T2DM), have a high risk for developing severe cases of COVID-19. The aim of this work was to characterize the alterations of blood immune cells (BIC) in patients with symptomatic COVID-19 and confirmed SARS-CoV-2 infection, ≥60 years and who needed hospitalization in the Centro de Salud Hospital of Tucuman, Argentina, during the second peak of the pandemic in Argentina. Ten patients were enrolled from December 2020 to May 2021. Blood samples were taken at the time of admission (day 0) and five days after (day 5) for routine laboratory tests and the characterization of BIC by flow cytometry. Most of the patients were men (70%) aged between 60 and 78 years. The 70% of patients had T2DM while 50% had arterial hypertension. At day 0, all the patients had increased neutrophils and inflammatory markers (C reactive protein and D-dimers) and reduced numbers of lymphocytes, HLA-DRhi monocytes, CD16+CD56+ NK cells, CD3+HLA−DR+CD25+ cells, CD4+ and CD8+ T cells in blood. Patients received a standard treatment for COVID-19 care (O2, corticosteroids and antibiotics). The hospital treatment normalized the levels of BIC (day 5) in 30% of patients who were those with no comorbidities. In patients with T2DM, BIC recovery was variable. In T2DM patients who required administration of plasma (30%), prolonged O2 therapy (40%) or referral to the intensive care unit (10%) significant reductions of CD16+CD56+, CD3+HLA−DR+CD25+, CD4+ and CD8+ cells were observed between days 0 and 5. In line with previous studies, our results show that absolute counts of major lymphocyte subsets in blood are significantly and substantially decreased during the course of severe COVID-19 disease in elderly patients. These BIC alterations may persist despite clinical care in elderly patients with T2DM. Further studies are needed to investigate the utility of early lymphocyte subset measurements as prognostic biomarkers of disease severity, mortality, and response to treatment in COVID-19 elderly patients with T2DM.

黏膜天疱疮患者循环性NC16-A特定T细胞的快速效应器功能

Background:Mucous membrane pemphigoid (MMP) is a chronic blistering skin dise ase frequently associated with circul atingautoantibodies directed to a number o f antigens including the NC16A region of BP180. NC16A domain-specific T cells h ave been identified in the blood of individuals with bullous pemphigoid (BP), pe mphigoid gestationis and linear IgA disease, but there are no data investigating the potential role for such T cells in the pathogenesis of MMP. Objectives:To test the hypothesis that NC16A-specific T cells exist in the peripheral blood o f individuals with MMP. Methods:We isolated peripheral blood mononuclear cells from 10 patients with MMP, 17 with BP and 10 healthy controls and examined the i mmunogenicity of overlapping peptides spanning the NC16A domain using interferon (IFN)-γenzyme-linked immunospot assay. Results:Significant IFN-γproductio n was observed in response to the NC16A peptides in two of the patients with MMP and two of the patients with BP but in none of the normal controls. These data suggest that in a minority of individuals with MMP, NC16A domain-specific T cel ls circulate at sufficiently high frequency to be detectable directly ex vivo an d to show rapid effector function. Conclusions:Overall, these findings are the first to examine the potential role for antigen-specific autoreactive T cells i n the pathogenesis of MMP, and confirm that in some individuals the NC16A domain may be an important target antigen.

Non Selective Inhibition of COX Activity Reversed Inflammation and Reactive Oxygen Radicals Mediated Prostate Cancer Risk and Decreased Disease Progression in Preclinical Model

Prostate cancer (PCa) represents the most frequent urologic diagnosis in elderly males. We have previously shown that exposure of prostate to lipopolysaccharide (LPS) promotes cancer risk. We investigated the effect of non-selective cyclooxygenase (COX) inhibition on prostate inflammation-mediated cancer risk in vivo. The prostates of male rats were inoculated with E. coli as sources of inflammatory molecules (LPS) and were treated with COX inhibitor, aspirin 2 mg/Kg orally for 14 days or PBS. Oxidative stress was induced with two 2 mls of hydrogen peroxide orally twice daily or PBS for 14 days;they were either treated with COX inhibitor or PBS for another 14 days. Blood was collected and analyzed for acid phosphatase and PSA. Data showed presences of LPS in the prostate of the rats resulted in gradual increase in PSA when compared to control (P < 0.0001). However, COX inhibition resulted in statistically significant reduction in concentration of PSA level compared to control group (P < 0.0001). To understand if oxidative stress mechanism was involved in the inflammation mediated increase in PSA, data showed that rats exposed to H2O2 had 2.5 fold increase in acid phosphatase (ACP) compared control (P < 0.0001), and by inhiting COX activity, a statistically significant reduction in ACP from 11.2 IU/L ± 0.67 to 5.7 IU/L ± 0.347 (P < 0.0034) was observed. Thus since increased in PSA was associated to cancer risk, our data suggested that inflammation mediated prostate cancer risk was reversible by Inhibition of COX Activity in rats.

Precision medicine in sepsis and septic shock:From omics to clinical tools

Sepsis is a heterogeneous disease with variable clinical course and several clinical phenotypes.As it is associated with an increased risk of death,patients with this condition are candidates for receipt of a very well-structured and protocolized treatment.All patients should receive the fundamental pillars of sepsis management,which are infection control,initial resuscitation,and multiorgan support.However,specific subgroups of patients may benefit from a personalized approach with interventions targeted towards specific pathophysiological mechanisms.Herein,we will review the framework for identifying subpopulations of patients with sepsis,septic shock,and multiorgan dysfunction who may benefit from specific therapies.Some of these approaches are still in the early stages of research,while others are already in routine use in clinical practice,but together will help in the effective generation and safe implementation of precision medicine in sepsis.

Human immunology and immunotherapy:main achievements and challenges

The immune system is a fascinating world of cells,soluble factors,interacting cells,and tissues,all of which are interconnected.The highly complex nature of the immune system makes it difficult to view it as a whole,but researchers are now trying to put all the pieces of the puzzle together to obtain a more complete picture.The development of new specialized equipment and immunological techniques,genetic approaches,animal models,and a long list of monoclonal antibodies,among many other factors,are improving our knowledge of this sophisticated system.The different types of cell subsets,soluble factors,membrane molecules,and cell functionalities are some aspects that we are starting to understand,together with their roles in health,aging,and illness.This knowledge is filling many of the gaps,and in some cases,it has led to changes in our previous assumptions;e.g.,adaptive immune cells were previously thought to be unique memory cells until trained innate immunity was observed,and several innate immune cells with features similar to those of cytokine-secreting T cells have been discovered.Moreover,we have improved our knowledge not only regarding immune-mediated illnesses and how the immune system works and interacts with other systems and components(such as the microbiome)but also in terms of ways to manipulate this system through immunotherapy.The development of different types of immunotherapies,including vaccines(prophylactic and therapeutic),and the use of pathogens,m onodonal antibodies,recombinant proteins,cytokines,and cellular immunotherapies,are changing the way in which we approach many diseases,especially cancer.

Beware the intruder:gasodermin A as molecular guardian preventing systemic dissemination of group A streptococci following local skin infection

Pyroptosis represents a host-protective mechanism that promotes clearance of pathogens by initiating the recruitment of immune cells to the site of infection.In a study recently published in Nature,Deng et al.extensively outlined a hitherto unknown role of the group A Streptococcus(GAS)-derived cysteine protease streptococcal pyrogenic exotoxin B(SpeB)in gasodermin A(GSDMA)cleavage as the primary step in induction of pyroptosis.As clearly shown in GSDMA-deficient mice infected with SpeBproducing GAS,this cleavage event is key to preventing the systemic spread and fatal course of GAS following local skin infection[1].